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Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3

The blood–brain barrier (BBB) greatly limits the delivery of protein-based drugs into the brain and is a major obstacle for the treatment of brain disorders. Targeting the transferrin receptor (TfR) is a strategy for transporting protein-based drugs into the brain, which can be utilized by using TfR...

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Autores principales: de la Rosa, Andrés, Metzendorf, Nicole G., Morrison, Jamie I., Faresjö, Rebecca, Rofo, Fadi, Petrovic, Alex, O’Callaghan, Paul, Syvänen, Stina, Hultqvist, Greta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9744743/
https://www.ncbi.nlm.nih.gov/pubmed/36509864
http://dx.doi.org/10.1038/s41598-022-25965-x
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author de la Rosa, Andrés
Metzendorf, Nicole G.
Morrison, Jamie I.
Faresjö, Rebecca
Rofo, Fadi
Petrovic, Alex
O’Callaghan, Paul
Syvänen, Stina
Hultqvist, Greta
author_facet de la Rosa, Andrés
Metzendorf, Nicole G.
Morrison, Jamie I.
Faresjö, Rebecca
Rofo, Fadi
Petrovic, Alex
O’Callaghan, Paul
Syvänen, Stina
Hultqvist, Greta
author_sort de la Rosa, Andrés
collection PubMed
description The blood–brain barrier (BBB) greatly limits the delivery of protein-based drugs into the brain and is a major obstacle for the treatment of brain disorders. Targeting the transferrin receptor (TfR) is a strategy for transporting protein-based drugs into the brain, which can be utilized by using TfR-binding BBB transporters, such as the TfR-binding antibody 8D3. In this current study, we investigated if binding to heparan sulfate (HS) contributes to the brain uptake of a single chain fragment variable of 8D3 (scFv8D3). We designed and produced a scFv8D3 mutant, engineered with additional HS binding sites, HS(+)scFv8D3, to assess whether increased HS binding would improve brain uptake. Additionally, a mutant with a reduced number of HS binding sites, HS(−)scFv8D3, was also engineered to see if reducing the HS binding sites could also affect brain uptake. Heparin column chromatography showed that only the HS(+)scFv8D3 mutant bound HS in the experimental conditions. Ex vivo results showed that the brain uptake was unaffected by the introduction or removal of HS binding sites, which indicates that scFv8D3 is not dependent on the HS binding sites for brain uptake. Conversely, introducing HS binding sites to scFv8D3 decreased its renal excretion while removing them had the opposite effect.
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spelling pubmed-97447432022-12-14 Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3 de la Rosa, Andrés Metzendorf, Nicole G. Morrison, Jamie I. Faresjö, Rebecca Rofo, Fadi Petrovic, Alex O’Callaghan, Paul Syvänen, Stina Hultqvist, Greta Sci Rep Article The blood–brain barrier (BBB) greatly limits the delivery of protein-based drugs into the brain and is a major obstacle for the treatment of brain disorders. Targeting the transferrin receptor (TfR) is a strategy for transporting protein-based drugs into the brain, which can be utilized by using TfR-binding BBB transporters, such as the TfR-binding antibody 8D3. In this current study, we investigated if binding to heparan sulfate (HS) contributes to the brain uptake of a single chain fragment variable of 8D3 (scFv8D3). We designed and produced a scFv8D3 mutant, engineered with additional HS binding sites, HS(+)scFv8D3, to assess whether increased HS binding would improve brain uptake. Additionally, a mutant with a reduced number of HS binding sites, HS(−)scFv8D3, was also engineered to see if reducing the HS binding sites could also affect brain uptake. Heparin column chromatography showed that only the HS(+)scFv8D3 mutant bound HS in the experimental conditions. Ex vivo results showed that the brain uptake was unaffected by the introduction or removal of HS binding sites, which indicates that scFv8D3 is not dependent on the HS binding sites for brain uptake. Conversely, introducing HS binding sites to scFv8D3 decreased its renal excretion while removing them had the opposite effect. Nature Publishing Group UK 2022-12-12 /pmc/articles/PMC9744743/ /pubmed/36509864 http://dx.doi.org/10.1038/s41598-022-25965-x Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
de la Rosa, Andrés
Metzendorf, Nicole G.
Morrison, Jamie I.
Faresjö, Rebecca
Rofo, Fadi
Petrovic, Alex
O’Callaghan, Paul
Syvänen, Stina
Hultqvist, Greta
Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title_full Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title_fullStr Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title_full_unstemmed Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title_short Introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scFv8D3
title_sort introducing or removing heparan sulfate binding sites does not alter brain uptake of the blood–brain barrier shuttle scfv8d3
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9744743/
https://www.ncbi.nlm.nih.gov/pubmed/36509864
http://dx.doi.org/10.1038/s41598-022-25965-x
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